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Enhancer of zeste acts as a major developmental regulator of Ciona intestinalis embryogenesis.

Le Goff E, Martinand-Mari C, Martin M, Feuillard J, Boublik Y, Godefroy N, Mangeat P, Baghdiguian S, Cavalli G - Biol Open (2015)

Bottom Line: These major phenotypic defects are specifically rescued by injection of a morpholino-resistant Ci-E(z) mRNA, which restores expression of Ci-E(z) protein and re-deposition of the H3K27me3 mark.As observed by qPCR analyses, Ci-E(z) invalidation leads to the early derepression of tissue-specific developmental genes, whereas late-acting developmental genes are generally down-regulated.Altogether, our results suggest that Ci-E(z) plays a major role during embryonic development in Ciona intestinalis by silencing early-acting developmental genes in a Hox-independent manner.

View Article: PubMed Central - PubMed

Affiliation: Université Montpellier, Place Eugène Bataillon, Montpellier 34095, Cedex 5, France Institut des Sciences de l'Evolution (ISEM), UMR5554, CNRS, Montpellier 34095, France.

No MeSH data available.


Related in: MedlinePlus

Localization of tissue-specific markers in Ci-E(z) morphants, control and rescue embryos. (A) Comparative in situ hybridization experiments were done with Ci-Fibrn (notochord specific) and Ci-MRLC2 (muscle specific) antisense probes at middle tailbud stage. MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; Rescue, mRNA-E(z)+MO-E(z) embryo. Number of embryos showing the presented expression pattern is indicated on each panel. For MRLC2, it is important to note that 27 of 27 morphants present muscle disorganization into the tail but only 12 of them have an additional ectopic expression in the head. (B) In situ hybridization negative controls done with Ci-Fibrn and Ci-MRLC2 sense probes. Scale bar: 30 µm.
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BIO010835F9: Localization of tissue-specific markers in Ci-E(z) morphants, control and rescue embryos. (A) Comparative in situ hybridization experiments were done with Ci-Fibrn (notochord specific) and Ci-MRLC2 (muscle specific) antisense probes at middle tailbud stage. MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; Rescue, mRNA-E(z)+MO-E(z) embryo. Number of embryos showing the presented expression pattern is indicated on each panel. For MRLC2, it is important to note that 27 of 27 morphants present muscle disorganization into the tail but only 12 of them have an additional ectopic expression in the head. (B) In situ hybridization negative controls done with Ci-Fibrn and Ci-MRLC2 sense probes. Scale bar: 30 µm.

Mentions: Finally the strong phenotype observed in Ci-E(z) morphants was characterized further by in situ hydridization using two specific markers respectively of notochord and striated muscle, namely probes specific of Fibrinogen-like (Fibrn) (Takahashi et al., 1999) and Myosin Regulatory Light Chain 2 (MRLC2) (Ikuta et al., 2010). As observed in Fig. 9, Fibrn expression was found to be mostly absent in Ci-E(z) morphants consistent with the lack of notochord formation previously described (Figs 2, 3, 4 and 6). MRLC2 however was found highly expressed in a massive bulk of cells densely packed but highly disorganized, reminiscent of the mis-positioning characterized at the ultrastructural level (Fig. 3). It is interesting to note that out of 27 analyzed Ci-E(z) morphants, 12 of them expressed MRLC2 ectopically in some anterior localized cells. Through rescue one could detect that both the expression of Fibrn and MRLC2 was partially and significantly corrected (20 out 22 embryos for Fibrn and 24 out of 28 for MRLC2) (Fig. 9).Fig. 9.


Enhancer of zeste acts as a major developmental regulator of Ciona intestinalis embryogenesis.

Le Goff E, Martinand-Mari C, Martin M, Feuillard J, Boublik Y, Godefroy N, Mangeat P, Baghdiguian S, Cavalli G - Biol Open (2015)

Localization of tissue-specific markers in Ci-E(z) morphants, control and rescue embryos. (A) Comparative in situ hybridization experiments were done with Ci-Fibrn (notochord specific) and Ci-MRLC2 (muscle specific) antisense probes at middle tailbud stage. MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; Rescue, mRNA-E(z)+MO-E(z) embryo. Number of embryos showing the presented expression pattern is indicated on each panel. For MRLC2, it is important to note that 27 of 27 morphants present muscle disorganization into the tail but only 12 of them have an additional ectopic expression in the head. (B) In situ hybridization negative controls done with Ci-Fibrn and Ci-MRLC2 sense probes. Scale bar: 30 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4582116&req=5

BIO010835F9: Localization of tissue-specific markers in Ci-E(z) morphants, control and rescue embryos. (A) Comparative in situ hybridization experiments were done with Ci-Fibrn (notochord specific) and Ci-MRLC2 (muscle specific) antisense probes at middle tailbud stage. MO-Ctrl, control embryo; MO-E(z), Ci-E(z) morphant; Rescue, mRNA-E(z)+MO-E(z) embryo. Number of embryos showing the presented expression pattern is indicated on each panel. For MRLC2, it is important to note that 27 of 27 morphants present muscle disorganization into the tail but only 12 of them have an additional ectopic expression in the head. (B) In situ hybridization negative controls done with Ci-Fibrn and Ci-MRLC2 sense probes. Scale bar: 30 µm.
Mentions: Finally the strong phenotype observed in Ci-E(z) morphants was characterized further by in situ hydridization using two specific markers respectively of notochord and striated muscle, namely probes specific of Fibrinogen-like (Fibrn) (Takahashi et al., 1999) and Myosin Regulatory Light Chain 2 (MRLC2) (Ikuta et al., 2010). As observed in Fig. 9, Fibrn expression was found to be mostly absent in Ci-E(z) morphants consistent with the lack of notochord formation previously described (Figs 2, 3, 4 and 6). MRLC2 however was found highly expressed in a massive bulk of cells densely packed but highly disorganized, reminiscent of the mis-positioning characterized at the ultrastructural level (Fig. 3). It is interesting to note that out of 27 analyzed Ci-E(z) morphants, 12 of them expressed MRLC2 ectopically in some anterior localized cells. Through rescue one could detect that both the expression of Fibrn and MRLC2 was partially and significantly corrected (20 out 22 embryos for Fibrn and 24 out of 28 for MRLC2) (Fig. 9).Fig. 9.

Bottom Line: These major phenotypic defects are specifically rescued by injection of a morpholino-resistant Ci-E(z) mRNA, which restores expression of Ci-E(z) protein and re-deposition of the H3K27me3 mark.As observed by qPCR analyses, Ci-E(z) invalidation leads to the early derepression of tissue-specific developmental genes, whereas late-acting developmental genes are generally down-regulated.Altogether, our results suggest that Ci-E(z) plays a major role during embryonic development in Ciona intestinalis by silencing early-acting developmental genes in a Hox-independent manner.

View Article: PubMed Central - PubMed

Affiliation: Université Montpellier, Place Eugène Bataillon, Montpellier 34095, Cedex 5, France Institut des Sciences de l'Evolution (ISEM), UMR5554, CNRS, Montpellier 34095, France.

No MeSH data available.


Related in: MedlinePlus